Competing roles of microRNAs and RNA-binding proteins in drug addiction

microRNA 和 RNA 结合蛋白在药物成瘾中的竞争作用

基本信息

批准号：
8657428
负责人：
Janet L Neisewander
金额：
$ 41.14万
依托单位：
UNIVERSITY OF NEW MEXICO HEALTH SCIS CTR
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-06-01 至 2017-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8657428
关键词：
3&apos Untranslated Regions Abstinence Addictive Behavior Animal Model Animal Testing Area Attention Attenuated BDNF gene Behavior Binding Binding Sites Brain Brain Diseases Brain region Brain-Derived Neurotrophic Factor Cells Cocaine Cocaine Abuse Cocaine Dependence Corpus striatum structure Crime Cues Databases Dimensions Disease Drug Addiction Drug abuse Drug usage Exhibits Extinction (Psychology)Functional RNA Gene Expression Gene Expression Regulation Gene Targeting Gene Transfer Genes Genetic Translation Goals Health Housing Illicit Drugs In Situ Hybridization In Vitro Infusion procedures Intake Intervention Justice Knowledge Measures Mediating Messenger RNA MicroRNAs Modeling Molecular Motivation Mus Neurons Neurosciences Neurosciences Research Nucleus Accumbens Outcome Pharmaceutical Preparations Play Post-Transcriptional Regulation Prosencephalon Protein Binding Psychological reinforcement RNA-Binding Proteins Rattus Regulation Repression Research Rewards Role Schedule Series Subfamily lentivirinae Synaptic plasticity Testing Time Training Transcript Viral Viral Vector Work addiction base cost drug relapse environmental enrichment for laboratory animals in vivo mRNA Stability mRNA Transcript Degradation neuron development novel overexpression preference productivity loss research study small hairpin RNA success tool

3&apos Untranslated Regions Abstinence Addictive Behavior Animal Model Animal Testing Area Attention Attenuated BDNF gene Behavior Binding Binding Sites Brain Brain Diseases Brain region Brain-Derived Neurotrophic Factor Cells Cocaine Cocaine Abuse Cocaine Dependence Corpus striatum structure Crime Cues Databases Dimensions Disease Drug Addiction Drug abuse Drug usage Exhibits Extinction (Psychology)Functional RNA Gene Expression Gene Expression Regulation Gene Targeting Gene Transfer Genes Genetic Translation Goals Health Housing Illicit Drugs In Situ Hybridization In Vitro Infusion procedures Intake Intervention Justice Knowledge Measures Mediating Messenger RNA MicroRNAs Modeling Molecular Motivation Mus Neurons Neurosciences Neurosciences Research Nucleus Accumbens Outcome Pharmaceutical Preparations Play Post-Transcriptional Regulation Prosencephalon Protein Binding Psychological reinforcement RNA-Binding Proteins Rattus Regulation Repression Research Rewards Role Schedule Series Subfamily lentivirinae Synaptic plasticity Testing Time Training Transcript Viral Viral Vector Work addiction base cost drug relapse environmental enrichment for laboratory animals in vivo mRNA Stability mRNA Transcript Degradation neuron development novel overexpression preference productivity loss research study small hairpin RNA success tool

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项目摘要

DESCRIPTION (provided by applicant): Controlling motivation for cocaine is the goal of long-term treatment success of cocaine addiction, which may require reversal of drug-induced changes in gene expression. Although post-transcriptional mechanisms play a vital role in the control of gene expression, their role in addictive behaviors has received little attention. RNA binding proteins and microRNAs serve as master switches controlling gene expression, with mRNA stability estimated to control about 20% of brain-expressed genes. Our research suggests that the RNA binding protein HuD and the microRNA miR-495 play opposite roles in the control of addiction-related gene expression and behavior: 1) They are predicted to bind the same GU-rich sequence in mRNAs; 2) Their binding sites are overrepresented in transcripts from an addiction-related gene (ARG) database; 3) They show differential regulation by cocaine in addiction-related brain regions, with miR-495 being downregulated and HuD upregulated; 3) In vitro manipulations of these molecules result in opposite effects on the expression of two of their target genes, BDNF and arc; 5) Most importantly, in vivo manipulations of these molecules show contrasting effects on motivation for cocaine. Based upon these results, we hypothesize that HuD and miR- 495 play a role in drug abuse by post-transcriptionally competing for binding to the same sequences and controlling the expression of ARGs in opposing directions. To test this hypothesis, we will: 1) test the functional competition between HuD and miR-495 for a) specific mRNA binding sites and the control of ARG gene expression, and b) cocaine conditioned place preference in mice overexpressing HuD in forebrain neurons; 2) determine the effects of viral-mediated gene transfer of miR- 495 and 3) HuD to the nucleus accumbens shell of rats using the following 3 models of motivation for cocaine: i) break point on a progressive ratio schedule of cocaine reinforcement, ii) extinction of cocaine-seeking behavior, and iii) reinstatement of extinguished cocaine-seeking behavior; 4) examine changes in the levels of miR-495 and HuD and selected target genes, including BDNF and arc in rats which have been manipulated to express varying degrees of motivation for cocaine. The proposed work synergistically combines the expertise of Dr. Perrone-Bizzozero in mRNA stability, HuD function, and target analyses and Dr. Neisewander in animal models of addiction and the neurocircuitry involved. The outcome of this work will provide new knowledge about the post-transcriptional mechanisms regulating addiction-related gene expression, an exciting new area of neuroscience research. A better understanding of these regulatory mechanisms is a pre-requisite for the application of these new tools in addiction research and ultimately in the treatment of this disorder.

描述（由申请人提供）：可卡因的控制动机是可卡因成瘾的长期治疗成功的目标，这可能需要逆转药物诱导的基因表达变化。尽管转录后机制在控制基因表达中起着至关重要的作用，但它们在成瘾行为中的作用很少受到关注。 RNA结合蛋白和microRNA充当控制基因表达的主开关，MRNA稳定性估计可控制约20％的脑表达基因。我们的研究表明，RNA结合蛋白HUD和microRNA miR-495在控制与成瘾相关的基因表达和行为中起相反的作用：1）它们被预测它们在mRNA中结合了相同的GU-rich序列； 2）它们的结合位点在与成瘾相关的基因（ARG）数据库中的转录本中过多代表； 3）它们显示可卡因在成瘾相关的大脑区域中对可卡因的差异调节，其中miR-495被下调和HUD上调。 3）对这些分子的体外操作会对其两个靶基因BDNF和ARC的表达产生相反的影响； 5）最重要的是，这些分子的体内操作对可卡因的动机显示出对比的影响。基于这些结果，我们假设HUD和miR-495通过转录后竞争与相同序列结合并控制ARG在相反方向上的表达来在药物滥用中发挥作用。为了检验这一假设，我们将：1）测试HUD和miR-495之间的功能竞争a）特定的mRNA结合位点和对ARG基因表达的控制，b）可卡因条件的位置偏好在前脑神经元中过表达HUD的小鼠中； 2) determine the effects of viral-mediated gene transfer of miR- 495 and 3) HuD to the nucleus accumbens shell of rats using the following 3 models of motivation for cocaine: i) break point on a progressive ratio schedule of cocaine reinforcement, ii) extinction of cocaine-seeking behavior, and iii) reinstatement of extinguished cocaine-seeking behavior; 4）检查miR-495和HUD的水平的变化以及选定的靶基因，包括大鼠中的BDNF和ARC，这些大鼠被操纵以表达可卡因的不同程度的动机。提出的工作协同结合了Perrone-Bizzozero博士在mRNA稳定性，HUD功能和目标分析中的专业知识，而Neisewander博士在成瘾的动物模型和涉及的神经回路中。这项工作的结果将提供有关调节成瘾相关基因表达的转录后机制的新知识，这是神经科学研究的令人兴奋的新领域。更好地理解这些调节机制是将这些新工具应用于成瘾研究并最终用于治疗这种疾病的先决条件。